Page 56 - 南京医科大学学报自然科学版

南京医科大学学报（自然科学版） 第41卷第9期
·1322 · Journal of Nanjing Medical University（Natural Sciences） 2021年9月

·基础研究·

WNT7b促进黑素瘤细胞的迁移、侵袭与上皮间质转化

陈奕鹤 ，马立文 ，殷旭峰 ，吴 迪 ，骆 丹 1*
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南京医科大学第一附属医院皮肤科，江苏 南京 210029；南京大学附属鼓楼医院皮肤科，江苏 南京 210008
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［摘 要］ 目的：明确WNT7b在恶性黑素瘤发生、发展中的作用，探索其促进恶性黑素瘤细胞发生侵袭转移的机制。方法：通
过 qRT⁃PCR 及 Western blot 检测人源黑素瘤细胞系中 WNT7b 的表达情况，挑选 WNT7b 高表达细胞系 A375，瞬时转染小干扰
RNA敲低WNT7b的表达。采用流式细胞仪分析细胞周期，划痕实验、Transwell 实验观察敲低WNT7b对A375细胞系增殖、迁
移和侵袭功能的影响。通过Western blot检测上皮⁃间质转化（epithelial⁃mesenchymal transition，EMT）关键标志物及与黑素瘤发
生发展密切相关的基质金属蛋白酶（matrix metalloproteinase，MMP）家族成员的表达。结果：WNT7b 在人恶性黑素瘤细胞系
A375、SK⁃MEL⁃28、A875中均有表达，A375的WNT76较其他细胞系表达量更高；敲低WNT7b使A375细胞生长阻滞于G1期，
同时细胞迁移与侵袭能力受到抑制。检测EMT相关蛋白，发现E⁃cadherin表达上调，N⁃cadherin和Vimentin表达下调。进一步
检测MMP家族成员的表达，发现MMP⁃2、MMP⁃7与MMP⁃9降低。结论：WNT7b在人源黑素瘤细胞中可能通过抑制MMP的表
达，诱导EMT的发生，从而促进黑素瘤细胞的迁移与侵袭。
［关键词］ 黑素瘤；WNT7b；上皮间质转化；基质金属蛋白酶
［中图分类号］ R739.6 ［文献标志码］ A ［文章编号］ 1007⁃4368（2021）09⁃1322⁃07
doi：10.7655/NYDXBNS20210908

WNT7b promotes migration，invasion and epithelial⁃mesenchymal transition of A375 cells

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CHEN Yihe ，MA Liwen ，YIN Xufeng ，WU Di ，LUO Dan 1*
1 Department of Dermatology，the First Affiliated Hospital of Nanjing Medical University，Nanjing 210029；
Department of Dermatology，Nanjing Drum Tower Hospital Affiliated to Nanjing University，Nanjing 210008，China
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［Abstract］ Objective：This study aims to determine the function of WNT7b in the progression of malignant melanoma and the
underlying mechanism by which it promotes the migration and invasion of melanoma cells. Methods：Intrinsic WNT7b expression level
was examined in A875，SK⁃MEL⁃28，and A375 cell lines by qRT⁃PCR and Western blot. The intervention of WNT7b in A375 cell line
was performed by transiently transfected with small interfering RNA，and its effect on the cellular function was also analyzed by flow
cytometry analysis，wound healing and Transwell assay，respectively. Then Western blot was used to detect the expression of epithelial⁃
mesenchymal transition（EMT）related markers and members of matrix metalloproteinase（MMP）family. Results：WNT7b was
expressed in A375，SK⁃MEL⁃28 and A875 cell lines，among which A375 presents the highest WNT7b expression. Our cellular study
showed that upon knockdown of WNT7b expression in human A375 cell line，the cell cycle was arrested at G1 phase，the cell migration
and invasion ability were significantly inhibited. In addition，WNT7b knockdown led to the up⁃regulation of E⁃cadherin and down⁃
regulation of N⁃cadherin，Vimentin and Snail1. Further detection of MMP family members showed that the expression of MMP⁃2，MMP⁃7
and MMP ⁃ 9 was reduced. Conclusion：WNT7b may induce EMT through MMPs in human malignant melanoma cells，thereby
promoting cells migration and invasion.
［Key words］ melanoma；WNT7b；epithelial⁃mesenchymal transition；matrix metalloproteinases
［J Nanjing Med Univ，2021，41（09）：1322⁃1328］

［基金项目］ 国家自然科学基金（81771512） 恶性黑素瘤是一种高度恶性的肿瘤，早期易发
∗ 生转移，近年来黑素瘤的发病率和病死率日益增
通信作者（Corresponding author），E⁃mail：daniluo2005@163.com